U.S. patent application number 16/545807 was filed with the patent office on 2020-06-11 for compositions, uses and methods for treating or preventing dental caries.
The applicant listed for this patent is APOLLONIA HEALTH INC.. Invention is credited to Dennis Gerard Cvitkovitch, Joon Kim.
Application Number | 20200179258 16/545807 |
Document ID | / |
Family ID | 50775341 |
Filed Date | 2020-06-11 |
United States Patent
Application |
20200179258 |
Kind Code |
A1 |
Kim; Joon ; et al. |
June 11, 2020 |
COMPOSITIONS, USES AND METHODS FOR TREATING OR PREVENTING DENTAL
CARIES
Abstract
The present invention provides compositions, uses and methods
thereof, for inhibiting the growth of caries-causing bacteria. The
composition comprises xylitol, sodium citrate, sodium bicarbonate,
anionic polymers and acceptable carrier materials. The compositions
can tend to be used for treating or preventing a condition caused
by caries-causing bacteria The compositions can further tend to be
used in patients such as children, adolescents or patients
suffering from a heightened susceptibility to toxic substances.
Inventors: |
Kim; Joon; (Hamilton,
CA) ; Cvitkovitch; Dennis Gerard; (Oakville,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
APOLLONIA HEALTH INC. |
HAMILTON |
|
CA |
|
|
Family ID: |
50775341 |
Appl. No.: |
16/545807 |
Filed: |
August 20, 2019 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14647380 |
May 26, 2015 |
|
|
|
PCT/CA2013/000978 |
Nov 25, 2013 |
|
|
|
16545807 |
|
|
|
|
61729804 |
Nov 26, 2012 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61Q 17/005 20130101;
A61K 8/733 20130101; A61K 2800/92 20130101; A61K 9/0095 20130101;
A61K 47/10 20130101; A61K 31/047 20130101; A61K 8/042 20130101;
A61K 2800/592 20130101; A61K 8/4993 20130101; A61K 31/194 20130101;
A61K 8/345 20130101; A61K 31/194 20130101; A61K 2800/30 20130101;
A61Q 11/00 20130101; A61K 33/00 20130101; A61K 2300/00 20130101;
A61K 8/022 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61P 1/02 20180101; A61K 33/00 20130101; A61K 8/365 20130101; A61P
31/04 20180101; A61K 47/36 20130101; A61K 31/047 20130101; A61K
8/362 20130101; A61K 8/19 20130101; A61K 8/60 20130101 |
International
Class: |
A61K 8/60 20060101
A61K008/60; A61Q 11/00 20060101 A61Q011/00; A61K 8/73 20060101
A61K008/73; A61K 8/49 20060101 A61K008/49; A61K 8/362 20060101
A61K008/362; A61K 8/34 20060101 A61K008/34; A61K 8/19 20060101
A61K008/19; A61K 31/047 20060101 A61K031/047; A61K 9/00 20060101
A61K009/00; A61K 47/10 20060101 A61K047/10; A61K 33/00 20060101
A61K033/00; A61K 47/36 20060101 A61K047/36; A61K 31/194 20060101
A61K031/194 |
Claims
1. A composition for inhibiting the growth of caries-causing
bacteria, comprising: sodium bicarbonate; sodium citrate; at least
one anionic polymer; and xylitol.
2. The composition of claim 1, wherein the sodium bicarbonate
concentration is between 0.1% to 1.5% w/w.
3. The composition of claim 1 or claim 2, wherein the sodium
citrate concentration is between 0.1% to 1% w/w.
4. The composition of claim 1, wherein the sodium bicarbonate to
sodium citrate ratio is between 1:1 to 2:1.
5. The composition of claim 1, wherein the sodium bicarbonate
concentration is 0.5% w/w.
6. The composition of claim 1, wherein the sodium citrate
concentration is 0.5% w/w.
7. The composition of claim 1, wherein the at least one anionic
polymer is sodium alginate.
8. The composition of claim 7 wherein the sodium alginate
concentration is between 0.05% to 0.5% w/w.
9. The composition of claim 8 wherein the sodium alginate
concentration is 0.1% w/w.
10. The composition of claim 1, wherein the xylitol concentration
is between 15% to 40% w/w.
11. The composition of claim 10 wherein the xylitol concentration
is 30% w/w.
12. The composition of claim 1 further comprising at least one
excipient.
13. The composition of claim 12 wherein the at least one excipient
is one or more of a binder, a lubricant, a disintegrant, a
suspending agent, an absorbent, a preservative, a surfactant, a
colorant, a suspending agent, water, glycerin, a flavouring agent,
an emulsifier, or polyglycitol syrup.
14. The composition of claim 13 wherein the water concentration is
between 35% to 85% w/w.
15. The composition of claim 14 wherein the water concentration is
between 50% to 75% w/w.
16. The composition of claim 15 wherein the water concentration is
58.75% w/w.
17. The composition of claim 13, wherein the glycerin concentration
is between 10% to 30% w/w.
18. The composition of claim 17 wherein the glycerin concentration
is between 15% to 25% w/w.
19. The composition of claim 17 wherein the glycerin concentration
is 10%.
20. The composition of claim 13, wherein the polyglycitol syrup
concentration is between 0% to 15% w/w.
21-43. (canceled)
Description
RELATED APPLICATIONS
[0001] This application claims priority from U.S. Provisional
Application No. 61/729,804 filed Nov. 26, 2013, the contents of
which are herein incorporated by reference.
FIELD OF INVENTION
[0002] This invention relates to compositions and methods for
treating or preventing dental caries. More particularly, the
present invention relates to compositions and methods for
inhibiting the growth of caries-causing bacteria.
BACKGROUND
[0003] Dental caries is an undesirable condition of the oral
cavity. Dental caries can cause extensive crown mutilations,
bacterial disorders of the periapical tissues, or even loss of the
affected dental elements. It is characterized by demineralization
of the dental enamel and dentin in various stages of progress until
it affects the pulp space. When the lesion passes beyond the
enamel-dentin border, a phlogistic reaction of the pulp tissues is
observed, with the formation of reaction dentin in some cases.
[0004] Dental caries is caused by bacteria that colonize in the
mouth and produce acid which eventually leads to dissolution of the
tooth enamel. These acidogenic bacteria include Streptococcus
mutans (S. mutans), Streptococcus sobrinus, and Lactobacili,
amongst others. These bacteria are resident of the biofilm
environment of dental plaque, a matrix of bacteria and
extracellular material that adheres to the tooth surface. Under
appropriate conditions, populations of these bacteria will rise and
the pH of the surrounding plaque will drop. These bacteria, being
among the most acid tolerant organisms residing in dental plaque,
will increase in numbers in this acidic environment and eventually
become a dominant member of the plaque community. Once attached,
the bacteria metabolize six-carbon dietary sugars, such as glucose,
sucrose, and fructose into lactic acid. This situation eventually
leads to dissolution of the tooth enamel, resulting in the
development of dental caries. Since there is a strong correlation
between the proportion of S. mutans in dental plaque or in saliva
relative to other bacterial species and the presence or risk of
future outbreaks of dental caries, S. mutans in plaque or saliva
may serve as an index for both dental caries activity state and
dental caries risk or susceptibility. These indices play an
increasingly important role in the diagnosis and treatment of
dental caries.
[0005] Recently it has become clear that dental plaque actually
consists of hundreds of different bacterial taxa. Most of these
bacteria exist on the surface of teeth in heterogeneous communities
called plaque or biofilms. The mouth thus acts as a reservoir for
these bacteria. While most of these bacteria are commensal, meaning
they fail to adversely affect the human host, others are pathogenic
and can cause tooth decay. Moreover these pathogenic bacteria have
been found to cause a life-threatening disease called
endocarditis.
[0006] S. mutans has also been found to contribute to infective
endocarditis. Infective endocarditis is a potentially lethal
infection of both native (normal) and artificial heart valves, and
if left untreated can be fatal. S. mutans forms biofilms on the
surface of these valves, and are typically a mixed community of a
variety of pathogenic bacteria. Since these bacteria find their way
to the valves via the blood stream and the blood stream is
typically aseptic, there are usually few opportunities for bacteria
to cause these infections. One exception is when pathogenic
bacteria, like S. mutans, enter the blood stream during dental
procedures. Hence, the probability of infection is directly related
to the reservoir of infectious bacteria found in the oral
cavity.
[0007] Previous efforts toward the correction of dental caries have
revolved around the use of the standard toothbrush to remove dental
plaque. Also in widespread use today are electric brushes, floss
and adjuncts such as proxy brushes. In addition, numerous
toothpastes and mouth rinses containing various supplements are
touted as aids in the prevention of dental caries. For example,
fluoride is commonly sold as a product for slowing the process of
dental decay. However, the efficacy of such methods of treating or
preventing dental caries is questionable. Dental plaque can only
partially be removed from the oral cavity, even when a demanding
regimen of oral hygiene that may include flossing, brushing and
regular visits to a dentist is followed. In addition, many
toothpastes and mouth rinses contain toxic supplements, such as
fluoride and triclosan, which can be toxic to very young
children.
[0008] In 2010, the US Surgeon General stated that dental caries is
the most common chronic disease in children; it is five times more
frequent than asthma and seven times more frequent than hay fever.
The National Health and Nutrition Examination Survey conducted from
1999 to 2004 found that close to one in two children between the
ages of 2 and 11 in the United States has at least one dental
cavity in his or her primary teeth. (See United States, National
Health and Nutrition Examination Survey, 1999-2004). The American
Academy of Pediatric Dentistry (AAPD) guidelines for reducing
caries incidence rates in infant and adolescent dental patients
recommends fluoridation in addition to periodic professional dental
services. Recommendations for pediatric fluoridation include a
"smear" amount of fluoridated toothpaste for children under the age
of 2 and a "pea-size" amount for children aged 2 to 5. In weighing
the risk-benefit profile of using fluoride in children, the AAPD
considered mild fluorosis (excessive fluorine consumption marked by
visible white spots on teeth) to be an insignificant health hazard
compared to prevention of the serious health risks of dental
disease (See American Academy Of Pediatric Dentistry, Guideline on
Infant Oral Health Care, 2012, Reference Manual V 35/NO 6
13/14).
[0009] Fluoride is also widely used in oral hygiene products. For
example, many toothpastes and mouth rinses contain fluoride. In
addition, many cities in Canada and the US have practiced public
water fluoridation for more than 60 years.
[0010] However, the systemic use of fluoride to prevent cavities is
controversial due to a potential correlation between cancer and
fluoride. According to The American Cancer Society, osteosarcoma
(cancer of the bone) is a rare form of cancer affecting 400
children and teens every year in the US. A number of long-term
animal and human study data has been collected, and in certain
studies, correlations have been found between higher levels of
fluoride in drinking water and elevated incidences of osteosarcoma
in both mice and humans, including children (See Levy M, Leclerc B
S. Fluoride in drinking water and osteosarcoma incidence rates in
the continental United States among children and adolescents.
Cancer Epidemiol. 2012:36: 83-88; See Comber H, Deady S, Montgomery
E, Gavin A. Drinking water fluoridation and osteosarcoma incidence
on the island of Ireland. Cancer Causes Control.
2011;22:919-924).
[0011] In its 2011 publication entitled Guideline on Xylitol Use in
Caries Prevention, the AAPD recommended using xylitol to prevent
caries in moderate to high-risk pediatric patients. The guideline
recommends administering xylitol dosages of between 3 and 8 grams
per day in divided doses to prevent caries in children of all ages.
Currently available oral hygiene products and inventions, however,
even if they do contain xylitol, tend to also include fluoride and
alcohol and other toxic substances. Accordingly these substances
can tend to be harmful when ingested, and thus particularly
unsuitable for dental caries prevention and treating compositions
for children and other vulnerable groups.
[0012] Despite many previous efforts to formulate a treatment to
treat or prevent dental caries, there is still a need for an
effective treatment for dental caries.
[0013] There is a need for improved oral compositions for
preventing and treating dental caries that do not contain fluoride.
There is also a need for providing oral compositions for preventing
and treating dental caries that are suitable for use by children
and other caries-vulnerable groups in the general population.
Further, there is a need for providing compositions that tend to
maintain a high pH in the oral cavity environment, as dental caries
causing bacteria thrive in acidic environments. Additionally, there
is a need for providing compositions for treating and preventing
dental caries that are longer lasting, and which do not contain
toxic substances that are harmful when ingested.
SUMMARY OF THE INVENTION
[0014] In an aspect of the present invention there is provided a
composition for inhibiting the growth of caries-causing bacteria,
comprising sodium bicarbonate, sodium citrate, at least one anionic
polymer and xylitol. In some embodiments the sodium bicarbonate
concentration is between 0.1% to 1.5% w/w, in other embodiments,
the sodium citrate concentration is between 0.1% to 1% w/w, and in
still further embodiments, either or both the sodium bicarbonate
and sodium citrate concentration is 0.5% w/w. In still further
embodiments, the sodium bicarbonate to sodium citrate ratio is
between 1:1 to 2:1.
[0015] In some embodiments the anionic polymer is sodium alginate,
and in further embodiments the sodium alginate concentration is
between 0.05% to 0.5% w/w, or is 0.1% w/w. In some embodiments the
xylitol concentration is between 15% to 40% w/w, or is 30% w/w.
[0016] In some aspects of the instant invention, the composition
further comprises at least one excipient. In some embodiments, the
excipient is a binder, a lubricant, a disintegrant, a suspending
agent, an absorbent, a preservative, a surfactant, a colorant, a
suspending agent, water, glycerin, a flavouring agent, an
emulsifier, or a polyglycitol syrup.
[0017] In some embodiments wherein the composition comprises water,
the water concentration can tend to be between 35% to 85% w/w,
between 50% to 75% w/w, or is 58.75% w/w. In some embodiments
wherein the composition comprises glycerin, the glycerin
concentration can tend to be between 10% to 30% w/w, between 15% to
25% w/w or is 10%. In some embodiments wherein the composition
comprises polyglycitol syrup, the polyglycitol concentration can
tend to be between 0% to 15% w/w or between 5% to 10% w/w.
[0018] In some aspects of the invention, the composition further
comprises at least one natural flavouring agent. The natural
flavouring agent concentration can tend to be less than 0.1% w/w or
can be 0.05% w/w. In some embodiments the natural flavouring agent
is a natural fruit flavor such as lemon.
[0019] In some aspects of the invention, the composition comprises
Polysorbate 20. In some embodiments wherein the composition
comprises Polysorbate 20, the Polysorbate 20 concentration can tend
to be between 0.01% to 0.1% w/w, or can be 0.1%.
[0020] In some aspects of the invention, the composition comprises
an anti-microbial agent. In some embodiments wherein the
composition comprises methylparaban as an anti-microbial agent, the
methylparaban concentration can tend to be less than 0.1% w/w.
[0021] In some aspects of the invention, the composition can tend
to be an oral formulation such as a liquid, a mouthwash, a
dentifrice, a varnish, a gel, a food product, a confectionary, an
ice cream, a chewing gum, a syrup, a cream, a tablet, a caplet, a
capsule, a chewable tablet, a quick dissolve tablet, an
effervescent tablet, a hard gelatin capsule, a soft gelatin
capsule, a powder, or a liquid suspension.
[0022] In another aspect of the present invention there is provided
a use of the composition for treating or preventing caries in a
patient. In some aspects the composition can be used by children,
adolescents or patient's suffering from a heightened susceptibility
to toxic substances.
[0023] In further embodiments of the present invention there are
provided uses of the composition for preparing a formulation for
treating or preventing caries, treating or preventing a condition
caused by caries-causing bacteria, preparing a formulation for
treating or preventing a condition caused by caries-causing
bacteria, inhibiting the growth of a caries-causing bacteria, and
preparing a formulation for inhibiting the growth of a
caries-causing bacteria. In some embodiments the caries-causing
bacteria are Streptococcus mutans, Streptococcus sobrinus, or
Lactobacili.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] For a better understanding of embodiments of the system and
methods described herein, and to show more clearly how they may be
carried into effect, reference will be made by way of example, to
the accompanying drawings in which:
[0025] FIG. 1 shows the pH of the culture medium after biofilms
were incubated with various test treatments for 1 minute,
[0026] FIG. 2 shows the drop in pH of the culture medium 6 hours
after biofilms were incubated with various test treatments for 1
minute.
[0027] FIG. 3 shows the drop in pH of the culture medium 8 hours
after biofilms were incubated with various test treatments for 1
minute.
[0028] FIG. 4 shows the survival of S. mutans cells 1 hour after a
1 minute incubation with various test treatments, wherein survival
was measured using a Presto Blue.TM. assay (*p<0.05).
[0029] FIG. 5 shows the survival of S. mutans cells 1 hour after a
1 minute incubation with various test treatments, wherein survival
was measured using a colony forming unit (CFU) assay
(*p<0.05).
DETAILED DESCRIPTION
[0030] The present invention has been described with regard to
specific embodiments. However, it will be obvious to persons
skilled in the art that a number of variants and modifications can
be made without departing from the scope of the invention as
described herein.
[0031] In particular, numerous specific details are set forth in
order to provide a thorough understanding of the embodiments
described herein. However, it will be understood by those of
ordinary skill in the art that the embodiments described herein may
be practiced without these specific details. In other instances,
well-known methods, procedures, and components have not been
described in detail so as not to obscure the embodiments described
herein. Furthermore, this description is not to be considered as
limiting the scope of the embodiments described herein in any way,
but rather as merely describing the implementation of the various
embodiments described herein.
[0032] The compositions for treating and preventing dental caries
described herein are particularly useful for administration to
children and other vulnerable groups. There are presently no
commercially available formulations for these consumers that are
specifically formulated to improve cavity prevention and treatment,
that include only Generally Recognized As Safe (GRAS) ingredients,
that increase pH for improved mineralization, and that increase
efficacy duration times.
[0033] As children tend to have weaker immune systems than adults,
parents generally avoid even small risks with respect to the health
of their children. The present invention tends to provide caries
prevention solutions to the most caries-vulnerable age group,
children aged 3 to 12, by increasing oral pH using bio-adhesive
materials to provide long lasting efficacy and safe ingredients.
Persons of skill will appreciate, however, that the compositions
described herein can be suitable for use by other populations,
include adult and geriatric populations.
[0034] In an embodiment of the present invention, there is provided
a composition for treating and preventing dental caries which tends
to increase the pH of the dental environment. Persons of skill will
appreciate that dental caries occur due to the production of acid
by bacteria that colonize in the mouth, leading to dissolution of
the tooth enamel. The pH balance of the inside of the mouth
determines how fast demineralization (the process of losing
mineral, mainly calcium, from tooth enamel) and remineralization
(the process of gaining mineral back to tooth enamel) occur to
determine the rate of caries occurrence in one's mouth. When the pH
range falls below 5.5, demineralization mainly occurs, and if the
low pH is maintained for a prolonged period of time, the cavity
occurrence increases drastically. Currently available oral hygiene
products, however, tend to have a low pH.
[0035] The present inventions provide hygiene products that
increase oral pH to promote remineralization and prevent
demineralization of tooth enamel. In some embodiments the
composition comprises sodium bicarbonate. In still further
embodiments, the composition comprises sodium citrate. These
ingredients tend to maintain a high pH in the oral cavity
environment. When these two salts are combined in an aqueous
solution, the anions, citrate and bicarbonate, act as alkalinizing
agents to increase the pH of the solution, thereby providing a
composition that tends to increase the pH of a dental environment
when present in the oral cavities of the mouth.
[0036] In some embodiments, Sodium bicarbonate content can range
between 0.1% to 1.5% w/w. In still further embodiments. Sodium
citrate can range between 0.1% to 1% w/w. In alternative
embodiments, the ratio of sodium bicarbonate and sodium citrate
tends to be between 1 to 1 and 2 to 1.
[0037] Whether the actives in mouthwash or toothpaste are
anti-bacterial or fluoride-like enamel protection agents, persons
of skill will appreciate that a general recommendation from dental
healthcare practitioners and manufacturers is to refrain from
eating or drinking for 30 minutes after rinsing or brushing with
standard cavity fighting agents. Eliminating the 30 minute waiting
time would be advantageous to the general population, including
active adults, busy parents and impatient children.
[0038] Embodiments of the present invention provide compositions
for controlled release of dental caries prevention and treatment
ingredients. In some embodiments, the compositions comprise anionic
polymers to absorb the active ingredients and then release the
active ingredients as hydration occurs. In further embodiments the
compositions comprise sodium alginate, naturally found in the cell
walls of brown algae, to control the release of other chemicals in
the composition. Such controlled release substances can tend to
reduce waiting times after application of the compositions
described herein.
[0039] In some embodiments, the natural bio-adhesive and
controlled-release material, sodium alginate, can tend to be used
to prolong the beneficial effects of composition ingredients, such
as xylitol, sodium bicarbonate and sodium citrate in the oral
cavity. Sodium alginate content can tend to be between 0.05% and
0.5% w/w.
[0040] Embodiments of the presently claimed compositions are
comprised of ingredients derived from natural sources or
ingredients that are Generally Recognized As Safe (GRAS) materials.
These safe ingredients can tend to be used in anticipation of
accidental ingestion by consumers, especially children.
[0041] Despite the lack of unequivocal confirmation that fluoride
is harmless, because of its known health benefits in caries
prevention, fluoride is widely used in oral hygiene products. Other
known formulations include water-soluble calcium salt to prevent or
treat cavities, although persons of skill will appreciate that the
use of calcium salt can tend to cause adverse effects. These
adverse effects include calcium overdose, especially if the
intended user group is children who drink homogenized milk or
consume calcium-rich food. Further known compositions for
preventing and treating caries tend to include alcohol, a substance
that acts as a well-known depressant when consumed.
[0042] Embodiments of the present invention tend not to contain
alcohol, fluoride or other chemicals deemed a risk when ingested.
The presently claimed invention provides effective caries
prevention without using harmful chemical agents that could pose
health risks to users if accidentally ingested.
[0043] Embodiments of the present invention comprise xylitol, a
sugar alcohol naturally found in many foods such as berries, oats,
and mushrooms. Xylitol can tend to reduce the ability of
caries-causing bacteria to adhere to the cell surface and to
inhibit the bacteria from metabolizing six-carbon dietary sugars
into lactic acid.
[0044] In a preferred embodiment, the composition can tend to
comprise between 3 to 8 grams of xylitol given in divided dosages.
The compositions described herein can tend to comprise between 15%
to 40% w/w xylitol, which tends to be required and tends to be
important to prevent cavities.
[0045] The compositions described herein can be made in a variety
of forms, such as the following compositions: mouthwash,
dentifrice, varnish, gel, confectionary, ice cream, chewing gum,
syrup, cream, and the like. In other embodiments, the compositions
can be made in other forms such as a tablet, a caplet, a capsule,
chewable tablet, a quick dissolve tablet, an effervescent tablet, a
hard gelatin capsule, a soft gelatin capsule, a powder, a liquid
suspension, and other types of food products. One skilled in the
art would recognize there are also other viable ways for delivering
the composition to a user. In an embodiment, the composition is in
a liquid dosage form; and in further embodiments, the composition
is in a mouthwash form.
[0046] Furthermore, these compositions can be made using
conventional equipment and techniques known in the art. When
preparing dosage forms incorporating the compositions of the
invention, the ingredients are normally blended with conventional
excipients such as binders, lubricants, disintegrants, suspending
agents, absorbents, preservatives, surfactants, colorants and
suspending agents.
[0047] Additional carrier materials can tend to be included, for
example water, glycerin, flavouring agents and emulsifyiers.
Persons of skill in the art will appreciate that other ingredients
can be included.
[0048] In embodiments of the present invention water can tend to be
present in the range of between 35% to 85% w/w. In further
embodiments, water can tend to be between 50% to 75% w/w. In
embodiments of the present invention wherein compositions are in
the form of mouthwash, water can tend to be the main solvent.
[0049] Glycerin, with or without polyglycitol syrup, can be used to
add sweetness, taste, viscosity and texture to embodiments of the
present invention. The range for glycerin content can tend to be
between 10% and 30% w/w. In further embodiments, the glycerin
content can tend to be between 15% and 25% w/w. The range for
polyglycitol syrup can tend to be between 0% and 15% w/w in some
embodiments, and between 5% and 10% w/w in further embodiments.
[0050] Natural fruit flavours, and not artificial flavours, can be
used in some embodiments to improve smell and taste of the
compositions. Natural flavouring agents can tend to be present at a
concentration of less than 0.1% w/w, possibly with a combination of
different flavours.
[0051] An emulsifier, Polysorbate 20, can tend to be included in
some embodiments to disperse the flavouring agents. For example,
the composition may contain between 0.01% to 0.1% w/w of
Polysorbate 20.
[0052] In some embodiments, the present invention is formulated to
prevent microbial contamination by regulating water activity (aw),
thereby reducing dependence on preservative agents. The
naturally-occurring anti-microbial agent, methyparaban, can be used
in some embodiments of the invention as a preservative to prevent
undesired microbial growth. In alternative embodiments, the
composition can comprise methylparaban in amounts of less than 0.1%
w/w.
[0053] Use of xylitol in combination with sodium citrate, sodium
bicarbonate and sodium alginate tends to provide a synergistic
effect. As discussed in the examples below, it has been observed
that in tests using S. mutans cultures, a composition containing
xylitol, sodium citrate, sodium bicarbonate and sodium alginate
results in an increase in the pH of the culture medium and a
decrease in S. mutans viability as compared with xylitol, or any of
the other active components, alone.
EXAMPLES
Example 1--Composition for inhibiting Dental Caries (Xyncal
8.3)
[0054] An example of an embodiment of a composition for inhibiting
dental caries, referred to throughout this application is:
TABLE-US-00001 Ingredient % w/w grams per 5 g dose Water 58.75
2.938 Glycerin 10.00 0.500 Xylitol 30.00 1.500 Sodium Bicarbonate
0.50 0.025 Sodium Citrate 0.50 0.025 Sodium Alginate 0.10 0.005
Flavor 0.05 0.003 Polysorbate 20 0.10 0.005
[0055] The composition of Example 1 will herein be referred to as
Xyncal 8.3.
Example 2--Potential Product Specification
[0056] A sample product specification for an embodiment of the
present invention is listed below:
TABLE-US-00002 Test Specification Solids, % w/w by hand
refractometer (.degree. brix) 25.0 +/- 2.0 pH, as is 08.3 +/- 0.3
Colour Colourless Appearance Clear liquid Odour Lemon Flavour
Sweet, lemon flavour Mouth feel and texture Slight body APC
(aerobic plate count). <20 Y&M (yeasts and moulds)
Example 3--Preparing S. mutans Biofilms
Bacteria Strain Used
[0057] Amongst acidogenic bacteria, S. mutans is the principal
etiological agent of caries in humans. Since there is a strong
correlation between the proportion of S. mutans in dental plaque or
in saliva relative to other bacterial species and the presence or
risk of future outbreaks of dental caries, S. mutans in plaque or
saliva may serve as an index for both dental caries activity state
and dental caries risk or susceptibility. S. mutans UA159 strain
was used in this Example.
Biofilm Preparation
[0058] On the first day, biofilm growth was initiated by
inoculating 10mL of 100% Todd-Hewitt broth (THYE) with 0.3% yeast
extract at a pH of 7.0 with UA159 to prepare overnight cultures.
The samples were incubated in closed screw-cap tubes without
agitation at 37.degree. C. in an anaerobic chamber (5% CO2),
[0059] On the second day, 1:100 dilutions were prepared by mixing
20 .mu.l of the overnight UA159 cultures with 1/4 THYE with 0.075%
yeast extract supplemented with 0.1% sucrose to reach a final
volume of 2 mL. The dilutions were then placed into the wells of
12-well plates. The cultures were incubated at 37.degree. C. for
18-24 hours without agitation in an anaerobic chamber (5% CO2).
Incubation with Test Treatments
[0060] On the third day, the 2 mL of medium was removed from each
well. 1.0 mL of test treatment solution, as described below, was
added to each well, after which the wells were incubated for 1
minute on the bench at room temperature. Next, the test treatment
solutions were removed, and 3.0 mL of undiluted THYE with 0.3%
yeast extract and no sucrose was added to each well. If an internal
pH control was required, fresh THYE was also added to two
un-inoculated wells. During these steps, care was taken to avoid
disrupting the biofilm.
[0061] The following test treatments were used:
TABLE-US-00003 Water Xyncal 8.3 Xylitol [1.5 g/5 ml] in water
Sodium bicarbonate [0.5 g/100 ml] in water Sodium citrate dihydrate
[0.5 g/100 ml] in water Sodium Alginate [0.1 g/100 ml] in water
Listerine .TM. Smart Rinse .TM. Listerine .TM. Agent Cool Blue
.TM.
Example 4--Effect of Composition of Example 1 (Xyncal 8.3) on pH in
Cultures Containing S. mutans
[0062] An in vitro assay was performed to determine whether the
compositions of the present invention would increase the pH in
culture media containing S. mutans as compared to control
treatments.
[0063] First, biofilms were prepared and incubated with test
treatments as described in Example 3. Immediately after 100% THYE
with 0.3% yeast extract was added to the wells following incubation
with the test treatments, the initial pH of the medium in each well
was measured. Samples were incubated at 37.degree. C. in an open
air, bench-top incubator, and the pH of the medium was measured
once an hour for the next 7-8 hours. Finally, the phi of the medium
in each well was measured again the next morning.
[0064] As shown in FIG. 1, a 1 minute incubation with Xyncal 8.3
resulted in the pH of the medium being above 5.5 for the duration
of the assay. Xyncal 8.3 was able to maintain a higher pH than any
of its individual components (xylitol, sodium citrate, sodium
bicarbonate or sodium alginate) on their own, exhibiting an
unexpected and synergistic effect. Listerine.TM. Smart Rinse.TM.,
however, was able to maintain the highest pH, which is believed to
be due to the inclusion of the antiseptic Cetylpyridinium Chloride
(CPC) in this rinse (which is also in Listerine.TM. Agent Cool
Blue.TM.). Since the rinse kills the biofilms, the pH of the medium
does not remain as low.
[0065] As compared to the water control, the results from Xyncal
8.3 showed a statistically significant difference at 1, 6, 7, 8,
and 24 hours (p<0.05); the results from Listerine.TM. Smart
Rinse.TM. showed a statistically significant difference at all
times (p<0.05); the results from xylitol showed a statistically
significant difference at 1, 6, 7, and 8 hours (p<0.05); the
results from sodium bicarbonate showed a statistically significant
difference at 1 hour (p<0.05); and the results from
Listerine.TM. Agent Cool Blue.TM. showed a statistically
significant difference at 1 to 8 hours (p<0.05). The results
from sodium citrate and sodium alginate showed no statistically
significant difference at any time.
[0066] FIGS. 2 and 3 shows more clearly the synergistic effect that
results when combining the components of Xyncal 8.3. FIG. 2 is a
graph showing the drop, or difference in total pH value, 6 hours
after incubation with the test treatments, i.e. the pH of the
medium at 6 hours subtracted from the initial pH of the medium,
wherein the test treatments are water, Xyncal 8.3. xylitol, sodium
bicarbonate, sodium citrate, and sodium alginate. FIG. 3 is a
similar graph showing the drop in pH 8 hours after treatment.
[0067] As can be seen from FIGS. 2 and 3, treating biofilms with
any one of sodium bicarbonate, sodium citrate, or sodium alginate
alone has little to no effect on the drop in pH; the drop is about
equal to that of the control, water. Of the components of Xyncal
8.3, only xylitol is able to slow the pH drop. One would therefore
expect that upon adding xylitol, sodium bicarbonate, sodium
citrate, and sodium alginate together, the pH drop would be about
as large as that for xylitol. However, as seen in FIGS. 2 and 3,
the pH drop of the medium when treated with Xyncal 8.3 is actually
lower than that for xylitol, showing synergy. Tables 1 to 8 below
show exemplary pH data obtained upon performing the in vitro assays
as described above.
TABLE-US-00004 TABLE 1 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Water Sample 1 7.32 6.9 6.48 5.8 4.86 4.68 4.68 4.67 4.75
Sample 2 7.34 6.91 6.69 6.48 6.09 5.57 5.04 4.93 Sample 3 7.26 6.88
6.73 6.56 6.26 5.76 5.21 4.93 4.94 Sample 4 7.54 7.04 6.89 6.61
6.29 5.79 5.23 4.97 4.91 Average 7.37 6.93 6.70 6.36 5.88 5.45 5.04
4.88 4.87 Standard 0.12 0.07 0.17 0.38 0.68 0.52 0.25 0.14 0.10
Deviation
TABLE-US-00005 TABLE 2 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Xyncal Sample 1 7.46 7.12 6.81 6.53 6.01 5.76 5.49 5.2
5.11 8.3 Sample 2 7.37 7.03 6.86 6.62 6.21 6 5.6 5.38 Sample 3 7.5
7.12 7.02 6.86 6.64 6.42 6.13 5.93 5.72 Sample 4 7.56 7.18 7.02
6.87 6.62 6.4 6.16 5.93 5.76 Average 7.47 7.11 6.93 6.72 6.37 6.15
5.85 5.61 5.53 Standard 0.08 0.06 0.11 0.17 0.31 0.32 0.35 0.38
0.36 Deviation
TABLE-US-00006 TABLE 3 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Listerine .TM. Sample 1 7.49 7.16 7.02 7.04 7.04 7.05 7
6.89 7.04 Smart Sample 2 7.34 7.02 6.89 6.83 6.8 6.91 6.81 6.78
Rinse .TM. Sample 3 7.59 7.13 7.01 6.97 6.92 6.98 6.92 6.9 6.89
Sample 4 7.57 7.13 7.03 6.99 6.95 6.97 6.96 6.93 6.92 Average 7.50
7.11 6.99 6.96 6.93 6.98 6.92 6.88 6.95 Standard 0.11 0.06 0.07
0.09 0.10 0.06 0.08 0.07 0.08 Deviation
TABLE-US-00007 TABLE 4 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Xylitol Sample 1 7.49 7.08 6.72 6.19 5.77 5.51 5.23 5.1
5.09 Sample 2 7.36 7 6.82 6.57 6.17 5.93 5.55 5.37 Sample 3 7.58
7.12 6.97 6.77 6.55 6.25 6.05 5.78 5.58 Sample 4 7.57 7.12 6.97 6.8
6.57 6.29 6.06 5.82 5.68 Average 7.50 7.08 6.87 6.58 6.27 6.00 5.72
5.52 5.45 Standard 0.10 0.06 0.12 0.28 0.38 0.36 0.41 0.34 0.32
Deviation
TABLE-US-00008 TABLE 5 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Sodium Sample 1 7.48 7.09 6.67 5.85 5.08 5.03 5 5 5.06
Bicar- Sample 2 7.39 6.97 6.81 6.6 6.21 5.7 5.11 4.96 bonate Sample
3 7.58 7.1 6.93 6.82 6.49 5.97 5.33 5.01 4.96 Sample 4 7.59 7.12
6.96 6.81 6.53 6.06 5.42 5.06 4.97 Average 7.51 7.07 6.84 6.52 6.08
5.69 5.22 5.01 5.00 Standard 0.09 0.07 0.13 0.46 0.68 0.47 0.19
0.04 0.06 Deviation
TABLE-US-00009 TABLE 6 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Sodium Sample 1 7.5 7.05 6.69 6.01 5.14 5.02 4.98 4.97
4.99 Citrate Sample 2 7.35 6.93 6.75 6.47 5.94 5.49 5.06 4.92
Sample 3 7.57 7.09 6.91 6.76 6.44 5.96 5.36 5.04 4.95 Sample 4 7.58
7.12 6.95 6.77 6.48 6.07 5.46 5.07 4.96 Average 7.50 7.05 6.83 6.50
6.00 5.64 5.22 5.00 4.97 Standard 0.11 0.08 0.12 0.36 0.62 0.48
0.23 0.07 0.02 Deviation
TABLE-US-00010 TABLE 7 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Sodium Sample 1 7.45 7.09 6.74 6.14 5.24 4.99 4.96 4.97
4.96 Alginate Sample 2 7.36 6.93 6.77 6.48 6 5.47 5.02 4.92 Sample
3 7.56 7.09 6.95 6.78 6.5 5.99 5.32 4.98 4.94 Sample 4 7.58 7.13
6.95 6.79 6.52 6.12 5.52 5.04 4.96 Average 7.49 7.06 6.85 6.55 6.07
5.64 5.21 4.98 4.95 Standard 0.10 0.09 0.11 0.31 0.60 0.52 0.26
0.05 0.01 Deviation
TABLE-US-00011 TABLE 8 test hours after treatment treatment 0 1 2 3
4 5 6 7 8 Listerine .TM. Sample 1 7.47 7.13 6.94 6.85 6.72 6.53 6.3
5.89 5.49 Agent Sample 2 7.38 7.02 6.89 6.8 6.74 6.77 6.63 6.51
Cool Sample 3 7.58 7.15 6.99 6.96 6.89 6.89 6.85 6.76 6.71 Blue
.TM. Sample 4 7.6 7.14 7 6.97 6.91 6.89 6.84 6.74 6.6 Average 7.51
7.11 6.96 6.90 6.82 6.77 6.66 6.48 6.27 Standard 0.10 0.06 0.05
0.08 0.10 0.17 0.26 0.41 0.67 Deviation
Example 5--Effect of Xyncal 8.3 on S. mutans Growth and Biofilm
Formation
[0068] An in vitro assay was performed to determine whether Xyncal
8.3 would show inhibitory effects on S. mutans growth and biofilm
formation.
[0069] First, biofilms were prepared and incubated with test
treatments as described in Example 3. immediately after undiluted
THYE with 0.3% yeast extract was added to the wells following
incubation with the test treatments, the samples were incubated for
1 hour at 37.degree. C.
[0070] Next, one of two different assays was performed to test S.
mutans growth and biofilm formation: a Presto Blue.TM. assay and a
Colony Forming Unit (CFU) assay.
[0071] For the Presto Blue.TM. assay, the biofilms were resuspended
in the media present in the wells. Next, the suspension was
sonicated for 20 seconds to disrupt bacterial chains. The
OD.sub.600 value was measured and adjusted to an OD.sub.600 value
of 0.2. A 90 .mu.L sample of the suspension was placed in the well
of a 96-well plate after which 10 .mu.L of Presto Blue.TM. reagent
was added to the well. The samples were incubated at 37.degree. C.
for at least 10 minutes or until the samples began to turn pink.
Finally, the samples were read in a plate reader at 570 nm and 600
nm and the 600 nm value was subtracted from the 570 nm value.
[0072] For the CFU assay, the biofilms were resuspended in the
media present in the wells. The suspensions were sonicated for 20
seconds to disrupt bacterial chains. Next, the suspensions were
serially diluted down to 10.sup.-7 by adding 20 .mu.L of the cell
suspension to 180 .mu.L of Phosphate Buffered Saline (PBS). For
each dilution, 20 .mu.L of solution was plated on 100% THYE agar
plates. After 24-48 hours, the CFUs were counted.
[0073] FIGS. 4 and 5 show the survival of S. mutans cells 1 hour
after a 1 minute incubation with the test treatments, measured
using the Presto Blue.TM. and CFU assays respectively. Both FIGS. 4
and 5 show that Xyncal 8.3 inhibits S. mutans growth and biofilm
formation to a greater extent than any of its individual components
(xylitol, sodium citrate, sodium bicarbonate or sodium alginate) on
their own, exhibiting an unexpected and synergistic effect.
[0074] The survival assays correlate with, and explain, the data
from Example 3. The increased inhibition of S. mutans growth and
biofilm formation with Xyncal 8.3, as compared with its individual
components (xylitol, sodiumitrate, sodium bicarbonate or sodium
alginate), explain why Xyncal 8.3 was able to maintain a higher pH
than any of its individual components. Listerine.TM. Smart
Rinse.TM.'s increased inhibition of S. mutans growth and biofilm
formation is believed to be due to the inclusion of the antiseptic
Cetylpyridinium Chloride (CPC) in this rinse (also in Listerine.TM.
Agent Cool Blue.TM.).
Example 6--Clinical Study
[0075] A statistically non-powered and non-randomized clinical
study was conducted involving eight consenting adults and one
teenager and one toddler under their parents' supervision.
[0076] Other than the toddler, who swallowed without much rinsing,
the participants rinsed their teeth once for 30 seconds with
between 2.5 ml to 5 ml the mouthwash composition from Example 1.
Nine participants chose to imbibe the mouthwash and one participant
spat it out after 30 seconds of gurgling.
[0077] All 10 participants enjoyed the flavour, odour, feel and
texture of the invention in the form of a mouthwash. No adverse
effect was reported after 24 hours, one week, one month, and three
months after imbibing or gurgling with the mouthwash.
[0078] Nine participants reported cleaner and more "pleasant"
feelings after rinsing their teeth with the mouthwash compared to
before rinsing, The "pleasant" feelings were described as cleaner,
sweeter, and even better than brushing with toothpaste. They also
reported that the "pleasant" feeling lasted between three hours and
12 hours. The toddler participant's parents reported that the
toddler's "morning breath" had significantly improved and smelt
pleasant in the morning.
[0079] The present invention has been described with regard to
specific embodiments. However, it will be obvious to persons
skilled in the art that a number of variants and modifications can
be made without departing from the scope of the invention as
described herein.
* * * * *